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Table 2 Summary of different approaches utilized for each genetic situations.

From: Determining the effectiveness of High Resolution Melting analysis for SNP genotyping and mutation scanning at the TP53 locus

Genetic situation HRM solutions
No common SNP in the amplicon Direct HRM analysis
Only one common SNP in the amplicon Sometimes it is possible to distinguish directly the three groups (Figure 1)
Otherwise, spike-in or unlabelled probe is needed (Figure 2)
Two SNPs in the amplicon, one is common the other is rare Spike-in when possible (Figure 3); otherwise, use unlabeled probe for the common SNP, and perform mutation scanning for the rare one (Figure 4)
Two or more common SNPs in the amplicon Use an unlabeled probe for each SNP (Figure 5)
Two SNPs in two different amplicons When Tm of 2 amplicons is different, it is possible to perform multiplex PCR (Figure 6)